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Overview

This clone has been shown to specifically recognise a fusion protein of PARK7/DJ1. It also recognises a FLAG-tagged PARK7/DJ1 expressed in eukaryotic cells.
A single band is seen when Western blotting in optimised conditions with this clone. However, overloading the gel or using low dilutions can cause other bands to appear.

Applications

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application

Abreviews

Notes

Flow Cyt

1/100. ab91545-Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.

IHC-Fr

Use at an assay dependent concentration.

ICC/IF

1/500.

WB

Use at an assay dependent concentration. Detects a band of approximately 20 kDa.

ICC

Use at an assay dependent concentration.

IHC-FoFr

1/1000.

Target

Function

Protects cells against oxidative stress and cell death. Plays a role in regulating expression or stability of the mitochondrial uncoupling proteins SLC25A14 and SLC25A27 in dopaminergic neurons of the substantia nigra pars compacta and attenuates the oxidative stress induced by calcium entry into the neurons via L-type channels during pacemaking. Eliminates hydrogen peroxide and protects cells against hydrogen peroxide-induced cell death. May act as an atypical peroxiredoxin-like peroxidase that scavenges hydrogen peroxide. Following removal of a C-terminal peptide, displays protease activity and enhanced cytoprotective action against oxidative stress-induced apoptosis. Stabilizes NFE2L2 by preventing its association with KEAP1 and its subsequent ubiquitination. Binds to OTUD7B and inhibits its deubiquitinating activity. Enhances RELA nuclear translocation. Binds to a number of mRNAs containing multiple copies of GG or CC motifs and partially inhibits their translation but dissociates following oxidative stress. Required for correct mitochondrial morphology and function and for autophagy of dysfunctional mitochondria. Regulates astrocyte inflammatory responses. Acts as a positive regulator of androgen receptor-dependent transcription. Prevents aggregation of SNCA. Plays a role in fertilization. Has no proteolytic activity. Has cell-growth promoting activity and transforming activity. May function as a redox-sensitive chaperone.

Defects in PARK7 are the cause of Parkinson disease type 7 (PARK7) [MIM:606324]. A neurodegenerative disorder characterized by resting tremor, postural tremor, bradykinesia, muscular rigidity, anxiety and psychotic episodes. PARK7 has onset before 40 years, slow progression and initial good response to levodopa. Some patients may show traits reminiscent of amyotrophic lateral sclerosis-parkinsonism/dementia complex (Guam disease).

Sequence similarities

Belongs to the peptidase C56 family.

Post-translationalmodifications

Sumoylated on Lys-130 by PIAS2 or PIAS4; which is enhanced after ultraviolet irradiation and essential for cell-growth promoting activity and transforming activity.Cys-106 is easily oxidized to sulfinic acid.Undergoes cleavage of a C-terminal peptide and subsequent activation of protease activity in response to oxidative stress.

Cellular localization

Cytoplasm. Nucleus. Mitochondrion. Under normal conditions, located predominantly in the cytoplasm and, to a lesser extent, in the nucleus and mitochondrion. Translocates to the mitochondrion and subsequently to the nucleus in response to oxidative stress and exerts an increased cytoprotective effect against oxidative damage. Detected in tau inclusions in brains from neurodegenerative disease patients.

ab11251 was shown to specifically react with PARK7/DJ1 in wild-type HAP1 cells. No band was observed when knockout samples were used. Wild-type and PARK7/DJ1 knockout samples were subjected to SDS-PAGE. ab11251 and ab181602 (loading control to GAPDH) were diluted at 1/500 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.

Overlay histogram showing HepG2 cells stained with ab11251 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab11251, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

I have investigated our records for ab11251. I am sorry toconfirm there is an error on the datsheet and that this antibody is sold as ascites. The datasheet has been updated with the correct information and I apol...

Thank you for your enquiry. Ab11251 has been tested for application in Western blotting and Immunocytochemistry, and has not yet been tested in any other applications (including IP). For Western blotting, I would recommend starting at a dilution of 1:5...

Thank you very much for your enquiry and for your patience. Ab11251 was originated outside Abcam and according to the originator, they had positive feedback from a researcher who found the antibody to cross-react with zebrafish. That is unfortunately a...